ADA preparation

All ADA procedures for this lab period was conducted by Puja Mody. The same approach was conducted during 09/19/2012

This was a two day procedure continued by Michael Nagle on the following day, 10/23/2012

The main concept for the procedure: E. coli was transformed to express ADA. After a 4 h. protein expression on an incubator shaker, the solution were centrifuge. The supernatant was collected and stored since this contained the soluble ADA while the pellets (cellular debris) were discarded.

Au/lysozyme solutions

The calculations were supposed to be posted on the notebook of Michael Nagle.

The calculation the author had executed for stock solution preparation is on 10/17/2012. The lysozyme stock solution was 15.24 μM and the gold was at 8.24 mM.

It was decided that the lysozyme would be kept at a 1.5 μM concentration for all sample varying the Au concentration. The same mole ratios from Au/BSA were used 60, 80, 100, 120, 128, 130, 132, 133, 134, 136, 138, 140, 160, and 170.

When the Au/lysozyme solutions were made, Nagle placed the rack in the Thermo Scientific incubator at 50°C for 4h.

AAS of Au/BSA solution of 10/17/12

The Au/BSA solutions were taken out from the oven. Mole ratios 60, 80, and 120 were the only solutions that exhibited a consistent, clear purple color with no fibers. This was indicative that all gold nanoparticles were in solution and none have nucleated to BSA.

Mole ratios 100 and others produced fibers, having solutions clear and colorless. The liquid was taken from each solution and transferred to falcon tubes; leaving some liquid suspending the fibers in the original tube. As a result, they were irregular amounts on each tube.

The irregular amounts created difficulty for centrifugation. Hours was spent in balancing the liquid for each pair of tubes.

Time went by and the fibers formed pellets on the bottom of the tubes. The formation of pellets was acceptable to allow to take AAS of the solutions.

The program for the Shimadzu AA-6200 Atomic Absorption Flame Emission was set on autozero. The flame was ignited exhibiting a blue-green color.

The blank used for the experiment was water.

The standards were run followed by an interval of sample-blank measurements. The calibration curve can be seen on the data section of Puja Mody's notebook.